Dominican University1, Chemistry, River Forest, IL 60305 Illinois State University2, Chemistry, Normal, IL 61790
Directed evolution is a powerful technique available to the beginning researcher. The application of standard techniques and equipment make it a valuable training tool for students. The use of multiple mutagenic techniques offers the student an applied lesson in molecular biology, and since the results are unknown, it is new every time. The experiment did not work as expected. The mutation, amplification, and ligation of the gene were optimized for the experimental setup. The transformation was substandard and the concentration of the expressed protein was too low for all quantification and qualification tests to be run. The use of SDS-PAGE to determine protein size as the screen for validating the genetic library was not adequate. DNA screening would be a better screening technique.
[Abstract (DOC)]